Bcl-2 Monoclonal Antibody (Bcl-2-100)

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	-Mouse (1:500)

Protocol tips
-Mouse (1:500)

Publication protocol

In brief, cells were seeded at a density of 4 × 105 cells in a 6 well plate and incubated for 24 h. Cells were subjected to treatments as described in the figure legends and processed for SDS-PAGE and western blotting as described previously63. In the present study, membrane was blocked with 5% non-fat milk in Tris buffered saline for 3 h at room temperature followed by overnight incubation with following primary antibodies including anti-FADD (1:1000), anti-cFLIPL (1:500), anti- p65 (1:1000), anti-ITCH (1:500), anti-PARP (1:1000), anti-cytochrome c (1:500), anti-caspase-3 (1:1000), anti-procaspase-8 (1:200), anti-procaspasse-7 (1:500), anti-procaspase-9 (1:1000), anti-JNK1 (1:200), anti-Bcl2 (1:500), anti-Ubiquitin (1:1000), anti-RIP1 (1:1000), anti-cIAP2 (1:250), anti-IKKβ (1:500) and anti-β-actin (1:2000) at 4 °C. After washing with TBST, the membrane was probed with horseradish peroxidase conjugated secondary antibodies (1:10,000). Expression of immunoreactive proteins was detected by using Novex® ECL HRP linked chemiluminiscent substrate kit (Invitrogen, USA) according to the instruction manual and developed in Kodak X-Omat blue film (NEN Life Sciences, Inc., Boston, MA) in the dark.

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Manufacturer protocol

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